The ventral hippocampus has emerged as a central hub for the orchestration of negative effects. We employed wireless amperometric O2 measurements in freely moving rats as a surrogate readout for changes in neuronal activity with translational value for fMRI studies in humans. From the Research Domain Criteria (RDoC) matrix, here, mainly one branch, namely the negative valence system is addressed using a modified Elevated Plus-Maze, male Wistar rats are tested under controllable (modified Elevated T-Maze, mETM with free exploration of one open and two closed arms) and uncontrollable (Open Arm Confinement, OAC) threatening conditions. Rats were equipped with an O2 sensor aimed at the right ventral CA1 region and treated with vehicle or midazolam (0.5mg/kg, intraperitoneal ( i.p)) 30 min before exposure to the mETM or OAC. On the mETM, midazolam not only exerted anxiolytic effects, but also reduced O2 signals. In case of the OAC, midazolam increased the escape rate while decreasing the O2 signals.Taken together, we qualified O2 measurements in defined threatening situations as a mean for linking anxiolytic pharmacological effects on behavior to concomitant changes in neuronal activity.